WO2024019542A1 - Liposome porphyrinique sensible aux ultrasons et son utilisation - Google Patents

Liposome porphyrinique sensible aux ultrasons et son utilisation Download PDF

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WO2024019542A1
WO2024019542A1 PCT/KR2023/010440 KR2023010440W WO2024019542A1 WO 2024019542 A1 WO2024019542 A1 WO 2024019542A1 KR 2023010440 W KR2023010440 W KR 2023010440W WO 2024019542 A1 WO2024019542 A1 WO 2024019542A1
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porphyrin
liposome
cancer
ultrasound
sensitive
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Korean (ko)
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정은아
김대현
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(주)아이엠지티
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0002—Galenical forms characterised by the drug release technique; Application systems commanded by energy
    • A61K9/0009—Galenical forms characterised by the drug release technique; Application systems commanded by energy involving or responsive to electricity, magnetism or acoustic waves; Galenical aspects of sonophoresis, iontophoresis, electroporation or electroosmosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
    • A61K41/0028—Disruption, e.g. by heat or ultrasounds, sonophysical or sonochemical activation, e.g. thermosensitive or heat-sensitive liposomes, disruption of calculi with a medicinal preparation and ultrasounds
    • A61K41/0033—Sonodynamic cancer therapy with sonochemically active agents or sonosensitizers, having their cytotoxic effects enhanced through application of ultrasounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/10—Dispersions; Emulsions
    • A61K9/127—Liposomes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/10—Dispersions; Emulsions
    • A61K9/127—Liposomes
    • A61K9/1271—Non-conventional liposomes, e.g. PEGylated liposomes, liposomes coated with polymers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
    • A61K41/0057—Photodynamic therapy with a photosensitizer, i.e. agent able to produce reactive oxygen species upon exposure to light or radiation, e.g. UV or visible light; photocleavage of nucleic acids with an agent
    • A61K41/0071—PDT with porphyrins having exactly 20 ring atoms, i.e. based on the non-expanded tetrapyrrolic ring system, e.g. bacteriochlorin, chlorin-e6, or phthalocyanines

Definitions

  • the present invention relates to ultrasound-sensitive liposomes containing porphyrin, methods for producing the same, and uses thereof.
  • cancer is still an important disease that ranks first and second among the causes of death worldwide.
  • Cancer treatment methods currently used clinically include chemotherapy, radiation, targeted therapy, and surgical resection of the lesion.
  • Representative examples of chemotherapy drugs for cancer treatment that have been used to date include Doxorubicin, Adriamycin, Cisplatin, Taxol, and 5-fluorouracil, and are widely used as chemotherapy for cancer treatment. It is being used extensively. However, the above methods have limitations and cause severe side effects and pain to patients rather than complete cure, so it is important to develop cancer treatment technologies that can minimize side effects.
  • existing anticancer drugs penetrate not only cancer cells but also normal tissues, damaging the function and activity of normal cells, causing side effects such as decreased bone marrow function, gastrointestinal disorders, and alopecia. It presents major problems in cancer treatment, such as multi-drug resistance. Therefore, research is being actively conducted on the development of innovative drug delivery systems that can reduce the non-specific effects of existing anticancer drugs and prevent unexpected side effects.
  • liposomes are vesicles composed of lipid bilayers surrounding an aqueous internal compartment, and the membrane is mainly composed of phospholipids and their derivatives.
  • the membrane is mainly composed of phospholipids and their derivatives.
  • phospholipids and their derivatives When phospholipids and their derivatives are dispersed in aqueous solutions, they spontaneously form endoplasmic reticulum.
  • liposomes can carry water-soluble active ingredients in the aqueous internal space and also can carry hydrophobic active ingredients in the lipid bilayer, they have been used as carriers for therapeutic agents such as drugs, enzymes, and gene sequences in the fields of medicine, pharmaceuticals, and biochemistry. .
  • liposomes developed to date have problems such as changes in the properties of the lipid bilayer due to the encapsulated drug, which impairs the structural stability of the liposome or its sensitivity to stimulation, and the pharmacological effect is halved due to non-specific drug release. There are problems with this or unexpected side effects occurring.
  • Porphyrin-liposomes containing porphyrin and phospholipids maintain high structural stability even when drugs are encapsulated, and can respond sensitively to ultrasound to release drugs. It was completed after confirming that it has the effect of improving drug delivery efficiency to target tissues and enhancing pharmacological activity.
  • the object of the present invention is to provide an ultrasound-sensitive porphyrin-liposome containing porphyrin and phospholipid.
  • Another object of the present invention is to provide a composition for drug delivery containing the ultrasound-sensitive porphyrin-liposome as an active ingredient.
  • Another object of the present invention is to provide a pharmaceutical composition for preventing or treating cancer, which contains the ultrasound-sensitive porphyrin-liposome encapsulated with an anticancer agent as an active ingredient.
  • Another object of the present invention is to provide a method for producing the ultrasound-sensitive porphyrin-liposome.
  • the porphyrin may generate reactive oxygen species when exposed to ultrasound, but is not limited thereto.
  • the ultrasound-sensitive porphyrin-liposome may have a lipid bilayer decomposed when exposed to ultrasound, but is not limited thereto.
  • the phospholipids include DOPE (1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine), DOPC (1,2-Dioleoyl-sn-glycero-3-phosphocholine), SOPC (1- Stearoyl-2-oleoyl-sn-glycero-3-phosphocholine), DEPC (1,2-Dierucoyl-sn-glycero-3-phosphocholine), POPE (1-palmitoyl-2-oleoyl- sn- glycero-3-phosphoethanolamine) , and POPG (1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol), but is not limited thereto.
  • DOPE 1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine
  • DOPC 1,2-Dioleoyl-sn-glycero-3-phosphocholine
  • SOPC 1- Stearoyl-2-oleoyl
  • the weight ratio of porphyrin:phospholipid may be 1:20 to 90, but is not limited thereto.
  • the porphyrin is contained at 1 to 10% by dry weight based on the total ultrasound-sensitive porphyrin-liposome, and the phospholipid is contained at 90 to 99% by dry weight based on the total ultrasound-sensitive porphyrin-liposome. It may be, but is not limited to this.
  • the ultrasound-sensitive porphyrin-liposome is DSPC (1,2-Distearoyl-sn-glycero-3-phosphorylcholine), DSPE-mPEG2000 (1,2-distearoyl-sn-glycero-3- It may further include one or more selected from the group consisting of phosphoethanolamine with conjugated methoxyl poly(ethylene glycol2000)), and cholesterol, but is not limited thereto.
  • the ultrasound-sensitive porphyrin-liposome may satisfy one or more characteristics selected from the group consisting of the following, but is not limited thereto:
  • (a) has a particle size of 100 to 200 nm
  • the ultrasound-sensitive porphyrin-liposome may be drug-encapsulated, but is not limited thereto.
  • the weight ratio of the ultrasound-sensitive porphyrin-liposome:drug may be 20 to 1:1, but is not limited thereto.
  • the present invention provides a drug delivery composition comprising the ultrasound-sensitive porphyrin-liposome as an active ingredient.
  • the present invention provides a drug delivery method comprising the step of administering the ultrasound-sensitive porphyrin-liposome to an individual in need thereof.
  • the ultrasound-sensitive porphyrin-liposome may be one in which the drug is encapsulated.
  • the present invention provides a drug delivery use of the ultrasound-sensitive porphyrin-liposome.
  • the present invention provides the use of the ultrasound-sensitive porphyrin-liposome for the production of a drug delivery system.
  • the ultrasound-sensitive porphyrin-liposome may be drug-encapsulated, but is not limited thereto.
  • the drug may be an anticancer agent, but is not limited thereto.
  • the present invention provides a pharmaceutical composition for the prevention or treatment of cancer, comprising the ultrasound-sensitive porphyrin-liposome as an active ingredient, wherein the ultrasound-sensitive porphyrin-liposome is encapsulated with an anticancer agent.
  • a pharmaceutical composition for treatment is provided.
  • the present invention provides a method for preventing or treating cancer, comprising the step of administering the ultrasound-sensitive porphyrin-liposome encapsulated with an anticancer agent to an individual in need thereof.
  • the present invention provides the use of the ultrasound-sensitive porphyrin-liposome encapsulated with an anticancer agent for the prevention or treatment of cancer.
  • the present invention provides the use of the ultrasound-sensitive porphyrin-liposome for the production of a drug for treating cancer (anticancer drug).
  • the anticancer agent is doxorubicin, paclitaxel, docetaxel, cisplatin, Gleevec, 5-fluorouracil (5-FU), tamoxifen, carboplatin, topotecan, belotecan, imatinib, irinotecan, and flocsuri.
  • doxorubicin doxorubicin
  • paclitaxel docetaxel
  • cisplatin Gleevec
  • 5-fluorouracil (5-FU) 5-fluorouracil
  • tamoxifen carboplatin
  • topotecan belotecan
  • imatinib irinotecan
  • flocsuri flocsuri.
  • It may be one or more selected from the group consisting of min, chlorambucil, busulfan, doxyfluridine, vinblastine, mitomycin, prednisone, Armitor, and mitoxantrone, but is not limited thereto.
  • the composition may be administered sequentially or simultaneously with ultrasonic treatment, but is not limited thereto.
  • the cancer is breast cancer, colon cancer, lung cancer, small cell lung cancer, stomach cancer, liver cancer, blood cancer, bone cancer, pancreatic cancer, skin cancer, head and neck cancer, cervical cancer, skin melanoma, intraocular melanoma, and uterine cancer.
  • the present invention includes the steps of (S1) dissolving porphyrin and phospholipid in a first organic solvent; and
  • step (S2) A method for producing an ultrasound-sensitive porphyrin-liposome, comprising the step of hydrating the mixed solution prepared in step (S1) in an aqueous solution and stirring it,
  • the first organic solvent may be one or more selected from the group consisting of dimethylacetamide, dimethylformamide, dimethyl sulfoxide, chloroform, methanol, ethanol, and ether, but is not limited thereto.
  • the phospholipids include DOPE (1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine), DOPC (1,2-Dioleoyl-sn-glycero-3-phosphocholine), SOPC (1-Stearoyl -2-oleoyl-sn-glycero-3-phosphocholine), DEPC (1,2-Dierucoyl-sn-glycero-3-phosphocholine), POPE (1-palmitoyl-2-oleoyl -sn- glycero-3-phosphoethanolamine), and POPG (1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol), but is not limited thereto.
  • DOPE 1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine
  • DOPC 1,2-Dioleoyl-sn-glycero-3-phosphocholine
  • SOPC 1-Stearoyl -2-oleoyl
  • step (S1) includes DSPC (1,2-Distearoyl-sn-glycero-3-phosphorylcholine), DSPE-mPEG2000 (1,2-distearoyl-sn-glycero) in addition to the porphyrin and phospholipid.
  • DSPC 1,2-Distearoyl-sn-glycero-3-phosphorylcholine
  • DSPE-mPEG2000 1,2-distearoyl-sn-glycero
  • cholesterol may be further dissolved in the first organic solvent, but is not limited thereto.
  • the present invention relates to ultrasound-sensitive porphyrin-liposomes capable of releasing drugs in response to ultrasound.
  • the ultrasound-sensitive porphyrin-liposome of the present invention is characterized in that it contains porphyrin. Porphyrin exposed to ultrasound reacts with water to generate reactive oxygen species (Reactive Oxygen Species) and constitutes a lipid bilayer of the porphyrin-liposome. Phospholipids react to reactive oxygen species generated by porphyrin, causing lipid peroxidation, which can eventually cause the structure of the lipid bilayer to collapse and release the encapsulated drug.
  • Reactive Oxygen Species reactive oxygen species
  • Phospholipids react to reactive oxygen species generated by porphyrin, causing lipid peroxidation, which can eventually cause the structure of the lipid bilayer to collapse and release the encapsulated drug.
  • the ultrasound-sensitive porphyrin-liposome according to the present invention can appropriately control drug release using ultrasound, and also maintains excellent structural stability even when the drug is encapsulated, thereby minimizing non-specific drug delivery.
  • the porphyrin-liposome of the present invention can improve the drug delivery efficiency and pharmacological effect of the drug, and can be used as an excellent drug carrier for various diseases by carrying various drugs.
  • Figure 1 is a TEM image of an ultrasound-sensitive porphyrin-liposome and a general liposome according to an embodiment of the present invention.
  • Figure 2 shows the results of confirming the drug release rate of drug-encapsulated ultrasound-sensitive porphyrin-liposomes or general liposomes depending on whether they were irradiated with ultrasound.
  • Figure 3 shows the results of confirming the cancer cell killing effect of ultrasound-sensitive porphyrin-liposomes and general liposomes each encapsulated with an anticancer agent, depending on whether or not they were irradiated with ultrasound.
  • Figure 4a shows the results of confirming the change in body weight over time after administering free doxorubicin or liposomes with or without ultrasound treatment to a cancer animal model.
  • Figure 4b shows the results of measuring tumor size over time after administering free doxorubicin or liposomes with or without ultrasound treatment to a cancer animal model.
  • Figure 5 is a diagram showing the action mechanism of the porphyrin-liposome according to the present invention with or without ultrasound.
  • the present invention relates to an ultrasound-sensitive porphyrin-liposome containing porphyrin and phospholipid. It was confirmed that the porphyrin-liposome maintains high structural stability even when encapsulated with a drug, and can respond sensitively to ultrasound to release the drug. It is complete. That is, the porphyrin-liposome according to the present invention can improve drug delivery efficiency to target tissues and enhance pharmacological activity, so it can be used as a drug carrier for various drugs.
  • the present invention provides an ultrasound-sensitive porphyrin-liposome comprising porphyrin and phospholipid.
  • the ultrasound-sensitive porphyrin-liposome may be briefly referred to as 'porphyrin-liposome'.
  • the porphyrin according to the present invention is characterized in that it reacts with water to generate reactive oxygen species (ROS) when exposed to sound waves, preferably ultrasonic waves. Therefore, porphyrin-liposomes containing porphyrin can generate reactive oxygen species due to porphyrin when exposed to ultrasound, and can result in structural changes due to phospholipids reacting to reactive oxygen species.
  • the porphyrin is a hydrophobic compound, and may be included (loaded, supported, or encapsulated) within the hydrophobic part of the porphyrin-liposome according to the present invention, such as a lipid bilayer membrane, but is not limited thereto.
  • the porphyrin according to the present invention contains iron (Fe) ions. More preferably, the porphyrin according to the present invention has 1 to 4, 1 to 3, 1 to 2, or 2 hydrogens (H) replaced with N-(2-(2-aminoethylamino)ethyl)butyramide. It could be. Most preferably, the porphyrin according to the present invention may be represented by the following formula (1), but is not limited thereto.
  • porphyrin of the present invention also includes pharmaceutically acceptable salts of porphyrin.
  • pharmaceutically acceptable salt includes salts derived from pharmaceutically acceptable inorganic acids, organic acids, or bases.
  • the term "pharmaceutically acceptable” means that the benefit/risk ratio is reasonable for use in contact with the tissue of a subject (e.g., a human) without undue toxicity, irritation, allergic reaction, or other problems or complications. It refers to a compound or composition that is suitable for the following and is within the scope of sound medical judgment.
  • acids examples include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, perchloric acid, fumaric acid, maleic acid, phosphoric acid, glycolic acid, lactic acid, salicylic acid, succinic acid, toluene-p-sulfonic acid, tartaric acid, acetic acid, citric acid, methanesulfonic acid, formic acid. , benzoic acid, malonic acid, gluconic acid, naphthalene-2-sulfonic acid, benzenesulfonic acid, etc.
  • Acid addition salts can be prepared by conventional methods, for example, by dissolving the compound in an excessive amount of aqueous acid and precipitating the salt using a water-miscible organic solvent such as methanol, ethanol, acetone, or acetonitrile. It can also be prepared by heating equimolar amounts of the compound and an acid or alcohol in water and then evaporating the mixture to dryness, or suction filtering the precipitated salt.
  • a water-miscible organic solvent such as methanol, ethanol, acetone, or acetonitrile.
  • Salts derived from suitable bases may include, but are not limited to, alkali metals such as sodium and potassium, alkaline earth metals such as magnesium, and ammonium.
  • the alkali metal or alkaline earth metal salt can be obtained, for example, by dissolving the compound in an excessive amount of alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the undissolved compound salt, and then evaporating and drying the filtrate.
  • an appropriate silver salt eg, silver nitrate
  • the scope of the compound of the present invention may include not only pharmaceutically acceptable salts, but also all isomers, hydrates, and solvates that can be prepared by conventional methods.
  • phospholipid refers to a complex lipid composed of glycerol, two fatty acid tails, and a phosphate-bonded head group.
  • Phospholipids are amphipathic compounds.
  • the hydrophilic head group reacts with the aqueous medium and the hydrophobic tail reacts with the organic solvent, so they are arranged at the interface between the aqueous medium and the organic solvent to spontaneously form a lipid bilayer. can be formed.
  • amphipathic compounds such as phospholipids are mixtures of two immiscible liquids (e.g. water and oil), mixtures of liquids and gases (e.g. gas microbubbles in water), or mixtures of liquids and insoluble particles (e.g. It is a compound that can stabilize mixtures of substances that are normally immiscible in other ways, such as metal nanoparticles in water.
  • the phospholipid according to the present invention is reactive to reactive oxygen species and is a phospholipid that can cause lipid peroxidation when exposed to reactive oxygen species.
  • the porphyrin-liposome according to the present invention is most preferably characterized by comprising one or more phospholipids containing unsaturated fatty acids.
  • porphyrin-liposome when the porphyrin-liposome according to the present invention is exposed to ultrasound, reactive oxygen species are generated by the porphyrin, resulting in lipid peroxidation of the porphyrin-liposome, ultimately destroying the lipid bilayer membrane of the porphyrin-liposome and releasing the encapsulated drug. It can be.
  • the phospholipid containing a carbon-carbon double bond is sufficient as long as it satisfies the above characteristics, and is not limited to specific types, but is preferably DOPE (1,2-Dioleoyl-sn-glycero-3- phosphoethanolamine), DOPC (1,2-Dioleoyl-sn-glycero-3-phosphocholine), SOPC (1-Stearoyl-2-oleoyl-sn-glycero-3-phosphocholine), DEPC (1,2-Dierucoyl-sn-glycero) -3-phosphocholine), POPE (1-palmitoyl-2-oleoyl- sn -glycero-3-phosphoethanolamine), and POPG (1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol).
  • DOPE 1,2-Dioleoyl-sn-glycero-3- phosphoethanolamine
  • DOPC 1,2-Dioleoyl
  • the porphyrin-liposome according to the present invention is characterized in that it does not contain specific phospholipids.
  • Specific fatty acids that are not included in the porphyrin-liposome according to the present invention include, for example, phosphatidyl choline (PC), dimyristoyl phospatidylcholine (DMPC), dipalmitoyl phosphatidycholine (DPPC), and egg It may be phosphatidyl glycerol (egg phosphatidyl glycerol), etc., but is not limited thereto.
  • the molar ratio (mole %) of total porphyrin:phospholipids constituting the porphyrin-liposome may be 1:500 to 2,000, but is not limited thereto.
  • the molar ratio of porphyrin:phospholipid is 1:500 to 1,800, 1:500 to 1,600, 1:800 to 2,000, 1:1,000 to 2,000, 1:1,200 to 2,000, 1:1,300 to 2,000, 1:1,400 to It may be 2,000, 1:1,400 to 1,800, or 1:1,400 to 1,600, but is not limited thereto.
  • the weight ratio of total porphyrin:phospholipids constituting the porphyrin-liposome may be 1:20 to 90, but is not limited thereto.
  • the weight ratio of the porphyrin:phospholipid is 1:20 to 90, 1:20 to 80, 1:20 to 70, 1:20 to 60, 1:20 to 50, 1:30 to 80, and 1:40. It may be 80, 1:30 to 70, 1:30 to 60, 1:40 to 60, or 1:40 to 50, but is not limited thereto.
  • the porphyrin may be included in an amount of 1 to 10% by dry weight based on the total porphyrin-liposome, but is not limited thereto. Specifically, the porphyrin may be included in a dry weight % of 1 to 10, 1 to 8, 1 to 6, 1 to 4, 1 to 3, or 1 to 2 based on the total porphyrin-liposome, but is not limited thereto.
  • the phospholipid may be included in an amount of 90 to 99% by dry weight relative to the total porphyrin-liposome, but is not limited thereto. Specifically, the phospholipid may be included at 90 to 99, 92 to 99, 94 to 99, 96 to 99, or 98 to 99% by dry weight based on the total porphyrin-liposome, but is not limited thereto.
  • porphyrin-liposome is DSPC (1,2-Distearoyl-sn-glycero-3-phosphorylcholine), DSPE-mPEG2000 (1,2-distearoyl-sn-glycero-3-phosphoethanolamine with conjugated methoxyl poly(ethylene) It may further include one or more selected from the group consisting of glycol2000)), and cholesterol.
  • the phospholipids according to the present invention are DOPE (1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine), DOPC (1,2-Dioleoyl-sn-glycero-3-phosphocholine), SOPC (1-Stearoyl-2- oleoyl-sn-glycero-3-phosphocholine), DEPC (1,2-Dierucoyl-sn-glycero-3-phosphocholine), POPE (1-palmitoyl-2-oleoyl- sn- glycero-3-phosphoethanolamine), POPG (1 -Palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol), DSPC (1,2-Distearoyl-sn-glycero-3-phosphorylcholine), DSPE-mPEG2000 (1,2-distearoyl-sn-glycero-3-phosphoethanolamine with conjugated methoxyl poly(ethylene glycol)
  • DSPC is an abbreviation for 1,2-Distearoyl-sn-glycero-3-phosphocholine (1,2-Distearoyl-sn-glycero-3-phosphocholine), and is a phosphatidylcholine head group (head group). refers to a phospholipid composed of two stearic acids attached to a group.
  • DSPE-mPEG2000 is 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-methoxy-poly(ethylene glycol-2000) (1,2-Distearoyl -sn-Glycero-3-Phosphoethanol amine with conjugated methoxyl poly(ethylene glycol2000)), refers to the pegylated derivative of 1,2-distearoyl-sn-glycero-3-PE (DSPE) .
  • cholesterol is a type of sterol (modified steroid) and refers to a lipid found in the cell membrane of all animal cells. Cholesterol according to the present invention includes derivatives of cholesterol. Derivatives of cholesterol may be, for example, cystosterol, ergosterol, stigmasterol, 4,22-stigmastadien-3-one, stigmasterol acetate, lanosterol, cycloartenol, or combinations thereof. Cholesterol is located in the lipid bilayer, and the permeability can be lowered or increased by controlling the amount, and can be used regardless of the ratio in the liposome.
  • the porphyrin-liposome further contains DSPC, DSPE-mPEG2000, and cholesterol
  • the DSPC:DSPE-mPEG2000:cholesterol:DOPE is in a weight ratio of 1:1 to 10:0.1 to 1:2 to 20. It may be included, but is not limited to this.
  • the DSPC:DSPE-mPEG2000:Cholesterol:DOPE is used at a weight ratio of 1:1 to 5:0.1 to 1:2 to 15, 1:1 to 10:0.1 to 1:2 to 10, 1:1 to 5:0.1 to 1:2 to 10, 1:1 to 5:0.1 to 0.9:2 to 10, 1:1 to 3:0.1 to 1:2 to 10, 1:1 to 3 : 0.5 to 1 : 2 to 8 by weight, 1 : 1 to 2 : 0.5 to 1 : 4 to 8, or 1 : 1 to 2 : 0.5 to 1 : 5 to 7, but limited thereto. It doesn't work.
  • the DSPC is 1 to 30% by weight relative to the total porphyrin-liposome
  • the DSPE-mPEG2000 is 5 to 40% by weight compared to the total porphyrin-liposome,
  • the cholesterol is 1 to 15% by weight relative to the total porphyrin-liposome, and
  • the DOPE may be included in an amount of 20 to 80% by weight based on the total porphyrin-liposome, but is not limited thereto.
  • the DSPC is 1 to 30% by weight, 1 to 25% by weight, 1 to 20% by weight, 1 to 15% by weight, 1 to 12% by weight, 5 to 30% by weight, 5 to 10% by weight, relative to the total porphyrin-liposome. It may be included at 20% by weight, 5 to 15% by weight, 7 to 12% by weight, 8 to 12% by weight, or 9 to 11% by weight, but is not limited thereto.
  • the DSPE-mPEG2000 is 5 to 40% by weight, 5 to 35% by weight, 5 to 30% by weight, 5 to 25% by weight, 5 to 20% by weight, 5 to 15% by weight, and 10 to 10% by weight, relative to the total porphyrin-liposome. It may be included at 40% by weight, 10 to 30% by weight, 10 to 25% by weight, 10 to 20% by weight, 15 to 30% by weight, 15 to 20% by weight, or 16 to 19% by weight, but is not limited thereto. .
  • the cholesterol is 1 to 15% by weight, 1 to 12% by weight, 1 to 10% by weight, 1 to 8% by weight, 5 to 15% by weight, 5 to 12% by weight, and 5 to 10% by weight relative to the total porphyrin-liposome. %, or 5 to 8 wt%, but is not limited thereto.
  • the DOPE is 20 to 80% by weight, 20 to 75% by weight, 20 to 70% by weight, 20 to 65% by weight, 30 to 80% by weight, 30 to 75% by weight, and 30 to 70% by weight relative to the total porphyrin-liposome.
  • % 30 to 65% by weight, 40 to 80% by weight, 40 to 75% by weight, 40 to 70% by weight, 40 to 65% by weight, 50 to 80% by weight, 50 to 75% by weight, 50 to 70% by weight, 50 to 65% by weight, 55 to 80% by weight, 55 to 75% by weight, 55 to 70% by weight, 55 to 65% by weight, 60 to 80% by weight, 60 to 75% by weight, 60 to 70% by weight, or 60% by weight. It may be included in an amount of 65% by weight, but is not limited thereto.
  • the DSPC:DSPE-mPEG2000:DOPE:cholesterol may be included in a molar ratio (mole%) of 1-50:1-80:5-80:0.1-50, but is not limited to this.
  • Liposomes satisfying the above lipid composition have been confirmed to have particularly excellent drug encapsulation rate, ultrasonic sensitivity, and stability in the blood.
  • the DSPC:DSPE-mPEG2000:DOPE:cholesterol has a molar ratio (mole%) of 1 ⁇ 50:1 ⁇ 80:5 ⁇ 80:0.1 ⁇ 50, 1 ⁇ 30:1 ⁇ 80:5 ⁇ 60:0.1 Molar ratio (mole%) of ⁇ 50, 1 ⁇ 30 : 1 ⁇ 60 : 5 ⁇ 60 : Molar ratio (mole%) of 0.1 ⁇ 30, molar ratio (mole%) of 1 ⁇ 30 : 1 ⁇ 60 : 5 ⁇ 60 : 0.1 ⁇ 10 %), 1 ⁇ 10 : 20 ⁇ 60 : 20 ⁇ 60 : 0.1 ⁇ 10 molar ratio (mole%), 1 ⁇ 15 : 30 ⁇ 50 : 30 ⁇ 50 : 1 ⁇ 5 molar ratio (mole%), 1 ⁇ 8 : 40 ⁇ 60 : 30 ⁇ 50 : molar ratio (mole%) of 1 ⁇ 10, 3 ⁇ 10 : 40 ⁇ 70 : 20 ⁇ 55 : molar ratio (mole%) of 0.1 ⁇ 3, 3 ⁇ 10 : 40 ⁇ 55 :
  • the DSPC is 1 to 20 mole ratio (mole%), 1 to 15 mole ratio (mole%), 1 to 10 mole ratio (mole%), 1 to 9 mole ratio (mole%), 1 to 8 mole ratio compared to the total porphyrin-liposome. (mole%), 1 to 7 mole ratio (mole%), 5 to 20 mole ratio (mole%), 5 to 15 mole ratio (mole%), 5 to 10 mole ratio (mole%), 5 to 9 mole ratio (mole%), It may be included in a mole ratio of 5 to 8 (mole%), or in a mole ratio of 5 to 7 (mole%), but is not limited thereto.
  • the DSPE-mPEG2000 has a molar ratio (mole%) of 10 to 80, a molar ratio (mole%) of 10 to 70, a molar ratio (mole%) of 10 to 65, and a molar ratio (mole%) of 10 to 60 compared to the total porphyrin-liposome.
  • molar ratio (mole%) from 10 to 55, molar ratio (mole%) from 10 to 50, molar ratio (mole%) from 10 to 48, molar ratio (mole%) from 20 to 80, molar ratio (mole%) from 20 to 70 ), molar ratio of 20 to 60 (mole%), molar ratio of 20 to 50 (mole%), molar ratio of 30 to 80 (mole%), molar ratio of 30 to 70 (mole%), molar ratio of 30 to 60 (mole%) ), a molar ratio (mole%) of 30 to 50, a molar ratio (mole%) of 30 to 48, a molar ratio (mole%) of 35 to 50, a molar ratio (mole%) of 40 to 50, or a molar ratio (mole%) of 45 to 50. %), but is not limited thereto.
  • the DOPE has a molar ratio (mole%) of 10 to 80, a molar ratio (mole%) of 10 to 70, a molar ratio (mole%) of 10 to 65, a molar ratio (mole%) of 10 to 60, and a molar ratio (mole%) of 10 to 55.
  • the cholesterol has a molar ratio (mole%) of 1 to 15, a molar ratio (mole%) of 1 to 10, a molar ratio (mole%) of 1 to 8, a molar ratio (mole%) of 1 to 6, and a molar ratio (mole%) of 1 to 5. (mole%), mole ratio of 1 to 4 (mole%), mole ratio of 2 to 10 (mole%), mole ratio of 2 to 8 (mole%), mole ratio of 2 to 5 (mole%), mole ratio of 2 to 4 (mole%), or may be included in a molar ratio (mole%) of 2 to 3, but is not limited thereto.
  • the porphyrin-liposome according to the present invention is an ultrasound-sensitive (ultrasound-sensitive) liposome that responds to ultrasound.
  • Ultrasound-sensitive liposome refers to a liposome whose permeability increases or its structure collapses when exposed to ultrasound. That is, when the porphyrin-liposome of the present invention is exposed to ultrasound, the lipid bilayer or structure of the porphyrin-liposome may collapse and the loaded drug may be released.
  • Ultrasound refers to a sound wave exceeding the frequency of 16 Hz to 20 kHz, which is the frequency of sound waves that the human ear can generally hear, and high-intensity focused ultrasound is a continuous, high-intensity sound wave.
  • focused ultrasonic waves that provide ultrasonic energy to a focal point, it is possible to produce instantaneous thermal effects (65-100°C cavitation effects, mechanical effects, and sonochemical effects) depending on energy and frequency.
  • Ultrasonic waves can produce effects on human tissue.
  • High-intensity ultrasound waves which are not harmful when passing through but form a focus, generate enough energy to cause coagulative necrosis and thermal cauterization effects regardless of the type of tissue.
  • the ultrasonic wave refers to a sound wave with a frequency higher than the audible frequency range of 16 Hz to 20 kHz.
  • the ultrasound may be high intensity focused ultrasound (HIFU), high intensity non-focused ultrasound, or a combination of the two, but is not limited thereto.
  • HIFU refers to ultrasound that focuses high-intensity ultrasound energy in one place and creates a concentrated focus.
  • ultrasound-guided high-intensity focused ultrasound Ultrasound-guided HIFU
  • MRI-guided HIFU magnetic resonance imaging-guided high-intensity focused ultrasound
  • the frequency of ultrasonic waves is, for example, 1 kHz to 100 kHz, 1 kHz to 90 kHz, 1 kHz to 80 kHz, 1 kHz to 70 kHz, 1 kHz to 60 kHz, 1 kHz to 50 kHz, 1 kHz to 40 kHz, 1 kHz to 30 kHz, 1 kHz to 20 kHz, 1 kHz to 10 kHz, 20 kHz to 3.0 MHz, 40 kHz to 2.0 MHz, 60 kHz to 2.0 MHz, 80 kHz to 2.0 MHz, 100 kHz to 2.0 MHz, 150 kHz to 2.0 MHz, 200 kHz to 2.0 MHz, 250 kHz to 2.0 MHz, 300 kHz to 2.0 MHz, 350 kHz to 2.0 MHz, 400 kHz to 2.0 MHz, 450 kHz to 2.0 MHz, 500 kHz to 2.0 MHz, 550 kHz to 2.0 MHz, 600 kHz to 2.0 MHz, 650 k
  • the intensity of ultrasonic waves is 0.1 to 30 W/cm 2 , 0.1 to 25 W/cm 2 , 0.1 to 20 W/cm 2 , 0.1 to 15 W/cm 2 , 0.1 to 10 W/cm 2 , 0.1 to 8 W/cm 2 , 0.1 to 5 W/cm 2 , 0.1 to 3 W/cm 2 , 0.1 to 2 W/cm 2 , 0.5 to 10 W/cm 2 , 0.5 to 8 W/cm 2 , 0.5 to It may be 5 W/cm 2 , 0.5 to 3 W/cm 2 , 0.5 to 2 W/cm 2 , 1 to 5 W/cm 2 , 1 to 3 W/cm 2 , or 1 to 2 W/cm 2 ; It is not limited to this.
  • the porphyrin-liposome according to the present invention may have guaranteed stability.
  • the above stability means that the drug encapsulated in the porphyrin-liposome is not released into the blood when not exposed to ultrasound.
  • the present inventors confirmed that the ultrasound-sensitive porphyrin-liposome according to the present invention maintains its structure stably even after a storage period of 3 months, and that almost no drug release occurs when not treated with ultrasound, so it has high structural stability. confirmed. In other words, the porphyrin-liposome according to the present invention maintains a stable structure in an environment without ultrasound, so almost no drug is released.
  • the particle size of the porphyrin-liposome according to the invention is, for example, 50 to 500 nm in diameter, 50 to 400 nm, 50 to 300 nm, 50 to 200 nm, 50 to 150 nm, 50 to 130 nm, 50 to 110 nm, 80 nm. It may be from 200 nm, 80 to 150 nm, 80 to 120 nm, 80 to 110 nm, 90 to 120 nm, or 90 to 120 nm, but is not limited thereto.
  • the particle size distribution of porphyrin-liposomes can be measured using Zetasizer Nano ZS (Malvern) using Dynamic Light Scattering (DLS) analysis after diluting the porphyrin-liposomes 10 times.
  • the method for measuring the particle size distribution of porphyrin-liposomes is not limited to this, and can be measured according to other methods known in the art and converted to equivalent values.
  • the ultrasound-sensitive porphyrin-liposome according to the present invention can carry a drug.
  • support may be used interchangeably with the terms “encapsulation” or “loading.”
  • the drug may be bound to the lipid bilayer membrane of the porphyrin-liposome, may be carried inside the heterobilayer, or may be carried in the water-soluble compartment inside the porphyrin-liposome.
  • the drug is supported inside the lipid bilayer of porphyrin-liposome.
  • the mixing ratio of porphyrin-liposome:drug is 20 to 1:1, 15 to 1:1, 10 to 1:1, 20 to 5:1, 15 to 5:1, 10 to 5:1. , 9 to 5:1, or 9 to 7:1 by weight (w/w%), but is not limited thereto, and any ratio that allows the drug to be efficiently encapsulated in the porphyrin-liposome is included without limitation.
  • the ratio of the drug loaded on the porphyrin-liposome compared to the total drug added may be 50 to 100%. That is, the porphyrin-liposome according to the present invention has a drug encapsulation ratio of 30% to 100%, 40% to 100%, 50% to 100%, 60% to 100%, 65% to 100%, and 70% to 100%. , 75% to 100%, 76% to 100%, 77% to 100%, 80% to 100%, 85% to 100%, 87% to 100%, 88% to 100%, 90% to 100%, or It may be 95% to 100%, but is not limited thereto.
  • the encapsulation rate may refer to the loading rate relative to the amount of added drug, but is not limited thereto.
  • the method of measuring the drug encapsulation rate of liposomes can be calculated by processing the drug into liposomes, separating the unencapsulated drug using size exclusive chromatography (SEC), and measuring the absorbance of the encapsulated and unencapsulated drugs. .
  • SEC size exclusive chromatography
  • the method for measuring the drug encapsulation rate is not limited to this, and can be measured according to other methods known in the art and converted to equivalent values.
  • the ultrasound-sensitive porphyrin-liposome encapsulated with the drug has 10% to 100%, 50% to 100%, 60% to 100%, 65% to 100%, 70% to 100%, Alternatively, it may have a drug release rate of 75% to 100%, but is not limited thereto.
  • the drug release rate may refer to the release rate relative to the amount of drug added, but is not limited thereto.
  • the release rate can be measured by treating the porphyrin-liposome with ultrasound, separating the drug released from the porphyrin-liposome and the porphyrin-liposome from which the drug was released by SEC, and measuring the absorbance of the drug.
  • the method for calculating the drug release rate is not limited to this, and can be measured according to other methods known in the art and converted to equivalent values.
  • the porphyrin-liposome encapsulated with the drug is 0.1% to 100%, 0.1% to 80%, 0.1% to 60%, 0.1% to 40%, 0.1% to 30%, It may have a drug release rate of 0.1% to 20%, 0.1% to 10%, or 30% or less, but is not limited thereto.
  • the present invention provides a drug delivery composition (i.e., drug delivery vehicle) comprising the ultrasound-sensitive porphyrin-liposome as an active ingredient.
  • a drug delivery composition i.e., drug delivery vehicle
  • the cells or tissues targeted for drug delivery are not limited to specific types, but are preferably cancer cells or tumor tissues.
  • the ultrasound-sensitive porphyrin-liposome may be one in which a drug is encapsulated.
  • the term “drug” refers to any compound that possesses the desired biological activity.
  • the biological activity of interest includes activity useful in the diagnosis, cure, alleviation, treatment, or prevention of disease in humans or other animals.
  • the drug may be an anticancer agent.
  • the present invention is a pharmaceutical composition for preventing or treating cancer, comprising the ultrasound-sensitive porphyrin-liposome as an active ingredient, wherein the ultrasound-sensitive porphyrin-liposome is characterized by encapsulating an anticancer agent. provides.
  • cancer or “tumor” as used in the present invention refers to aggressive characteristics in which cells divide and grow in defiance of normal growth limits, invasive characteristics that infiltrate surrounding tissues, It is a general term for diseases caused by cells with metastatic characteristics that spread to other parts of the body.
  • the type of cancer is not particularly limited as long as it is a malignant tumor known in the art, and includes, for example, breast cancer, colon cancer, lung cancer, small cell lung cancer, stomach cancer, liver cancer, blood cancer, bone cancer, pancreatic cancer, skin cancer, and head and neck cancer.
  • cervical cancer skin melanoma, intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, anal cancer, colon cancer, fallopian tube carcinoma, endometrial carcinoma, cervical cancer, vaginal cancer, vulvar carcinoma, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine cancer.
  • thyroid cancer parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, leukemia, lymphocytic lymphoma, bladder cancer, kidney cancer, ureteral cancer, renal cell carcinoma, renal pelvic carcinoma, CNS tumor, primary CNS lymphoma, may be selected from the group consisting of spinal cord tumor, brainstem glioma, and pituitary adenoma.
  • anticancer agent is not particularly limited as long as it can exert an anticancer effect, such as inhibiting tumor growth or metastasis of cancer.
  • the content of the drug-encapsulated porphyrin-liposome in the composition of the present invention can be appropriately adjusted depending on the symptoms of the disease, the degree of progression of the symptoms, the patient's condition, etc., for example, 0.0001 to 99.9% by weight based on the total weight of the composition, or It may be 0.001 to 50% by weight, but is not limited thereto.
  • the content ratio is a value based on the dry amount with the solvent removed.
  • the pharmaceutical composition according to the present invention may further include appropriate carriers, excipients, and diluents commonly used in the preparation of pharmaceutical compositions.
  • the excipient may be, for example, one or more selected from the group consisting of diluents, binders, disintegrants, lubricants, adsorbents, humectants, film-coating materials, and controlled-release additives.
  • the pharmaceutical composition according to the present invention can be prepared as powder, granules, sustained-release granules, enteric-coated granules, solutions, eye drops, ellipsis, emulsions, suspensions, spirits, troches, perfumes, and limonadese according to conventional methods.
  • Carriers, excipients, and diluents that may be included in the pharmaceutical composition according to the present invention include lactose, dextrose, sucrose, oligosaccharides, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, and calcium. These include phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.
  • diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants.
  • Additives to tablets, powders, granules, capsules, pills, and troches according to the present invention include corn starch, potato starch, wheat starch, lactose, white sugar, glucose, fructose, di-mannitol, precipitated calcium carbonate, synthetic aluminum silicate, and phosphoric acid.
  • Excipients such as cellulose (HPMC) 1928, HPMC 2208, HPMC 2906, HPMC 2910, propylene glycol, casein, calcium lactate, and Primogel; Gelatin, gum arabic, ethanol, agar powder, cellulose acetate phthalate, carboxymethyl cellulose, calcium carboxymethyl cellulose, glucose, purified water, sodium caseinate, glycerin, stearic acid, sodium carboxymethyl cellulose, sodium methyl cellulose, methyl cellulose, microcrystalline cellulose, dextrin.
  • binders can be used, Hydroxypropyl methyl cellulose, corn starch, agar powder, methyl cellulose, bentonite, hydroxypropyl starch, sodium carboxymethyl cellulose, sodium alginate, calcium carboxymethyl cellulose, calcium citrate, sodium lauryl sulfate, silicic acid anhydride, 1-hydroxy Propylcellulose, dextran, ion exchange resin, polyvinyl acetate, formaldehyde-treated casein and gelatin, alginic acid, amylose, guar gum, sodium bicarbonate, polyvinylpyrrolidone, calcium phosphate, gelled starch, gum arabic, Disintegrants such as amylopectin, pectin, sodium polyphosphate, ethyl
  • soybean oil (Lubri wax), aluminum stearate, zinc stearate, sodium lauryl sulfate, magnesium oxide, Macrogol, synthetic aluminum silicate, silicic anhydride, higher fatty acids, higher alcohol, silicone oil, paraffin oil, polyethylene glycol fatty acid ether, Lubricants such as starch, sodium chloride, sodium acetate, sodium oleate, dl-leucine, and light anhydrous silicic acid may be used.
  • Additives for the liquid according to the present invention include water, dilute hydrochloric acid, dilute sulfuric acid, sodium citrate, sucrose monostearate, polyoxyethylene sorbitol fatty acid esters (twin esters), polyoxyethylene monoalkyl ethers, lanolin ethers, Lanolin esters, acetic acid, hydrochloric acid, aqueous ammonia, ammonium carbonate, potassium hydroxide, sodium hydroxide, prolamine, polyvinylpyrrolidone, ethyl cellulose, sodium carboxymethyl cellulose, etc. can be used.
  • a solution of white sugar, other sugars, or sweeteners, etc. may be used in the syrup according to the present invention, and if necessary, flavoring agents, colorants, preservatives, stabilizers, suspending agents, emulsifiers, thickening agents, etc. may be used.
  • Purified water can be used in the emulsion according to the present invention, and emulsifiers, preservatives, stabilizers, fragrances, etc. can be used as needed.
  • Suspensions according to the present invention include acacia, tragacantha, methylcellulose, carboxymethylcellulose, sodium carboxymethylcellulose, microcrystalline cellulose, sodium alginate, hydroxypropylmethylcellulose (HPMC), HPMC 1828, HPMC 2906, HPMC 2910, etc. Topics may be used, and surfactants, preservatives, stabilizers, colorants, and fragrances may be used as needed.
  • Injections according to the present invention include distilled water for injection, 0.9% sodium chloride injection, IV solution, dextrose injection, dextrose + sodium chloride injection, PEG, lactated IV solution, ethanol, propylene glycol, non-volatile oil - sesame oil.
  • solvents such as cottonseed oil, peanut oil, soybean oil, corn oil, ethyl oleate, isopropyl myristic acid, and benzene benzoate
  • Solubilizers such as sodium benzoate, sodium salicylate, sodium acetate, urea, urethane, monoethylacetamide, butazolidine, propylene glycol, Tween, nicotinic acid amide, hexamine, and dimethylacetamide
  • Weak acids and their salts acetic acid and sodium acetate), weak bases and their salts (ammonia and ammonium acetate), organic compounds, proteins, albumin, peptone, and buffering agents such as gums
  • Isotonic agents such as sodium chloride
  • Stabilizers such as sodium bisulfite (NaHSO 3 ) carbon dioxide gas, sodium metabisulfite (Na 2 S 2 O 5 ), sodium sulfite (Na 2 SO 3 ), nitrogen gas (N 2 ),
  • Suppositories according to the present invention include cacao oil, lanolin, witepsol, polyethylene glycol, glycerogelatin, methylcellulose, carboxymethylcellulose, a mixture of stearic acid and oleic acid, Subanal, cottonseed oil, peanut oil, palm oil, cacao butter + Cholesterol, lecithin, Lanet wax, glycerol monostearate, Tween or Span, Imhausen, monolene (propylene glycol monostearate), glycerin, Adeps solidus, Buytyrum Tego -G), Cebes Pharma 16, Hexalide Base 95, Cotomar, Hydrocote SP, S-70-XXA, S-70-XX75(S-70-XX95), Hydro Hydrokote 25, Hydrokote 711, Idropostal, Massa estrarium (A, AS, B, C, D, E, I, T), Massa-MF, Massaupol, Masupol-15, Neosupostal-
  • Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations include the extract with at least one excipient, such as starch, calcium carbonate, and sucrose. ) or prepared by mixing lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium styrate talc are also used.
  • Liquid preparations for oral administration include suspensions, oral solutions, emulsions, and syrups.
  • various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included.
  • Preparations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories.
  • Non-aqueous solvents and suspensions include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate.
  • composition according to the present invention is administered in a pharmaceutically effective amount.
  • pharmaceutically effective amount means an amount sufficient to treat the disease with a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is determined by the type, severity, drug activity, and It can be determined based on factors including sensitivity to the drug, time of administration, route of administration and excretion rate, duration of treatment, drugs used simultaneously, and other factors well known in the medical field.
  • the pharmaceutical composition according to the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or multiple times. Considering all of the above factors, it is important to administer an amount that can achieve the maximum effect with the minimum amount without side effects, and this can be easily determined by a person skilled in the art to which the present invention pertains.
  • the pharmaceutical composition of the present invention can be administered to an individual through various routes. All modes of administration are contemplated, including oral administration, subcutaneous injection, intraperitoneal administration, intravenous injection, intramuscular injection, paraspinal space (intrathecal) injection, sublingual administration, buccal administration, intrarectal injection, vaginal injection. It can be administered by internal insertion, ocular administration, ear administration, nasal administration, inhalation, spraying through the mouth or nose, dermal administration, transdermal administration, etc.
  • the pharmaceutical composition of the present invention is determined depending on the type of drug as the active ingredient along with various related factors such as the disease to be treated, the route of administration, the patient's age, gender, weight, and severity of the disease.
  • the effective amount of the composition according to the present invention may vary depending on the patient's age, gender, and body weight, and is generally administered at 0.001 to 150 mg, preferably 0.01 to 100 mg, per kg of body weight every day or every other day, or 1 It can be administered in divided doses 1 to 3 times a day.
  • the above dosage does not limit the scope of the present invention in any way.
  • “individual” refers to a subject in need of treatment for a disease, and more specifically, human or non-human primates, mice, rats, dogs, cats, horses, cows, etc. refers to mammals of
  • “administration” means providing a given composition of the present invention to an individual by any suitable method.
  • prevention refers to any action that suppresses or delays the onset of the desired disease
  • treatment refers to the improvement or improvement of the desired disease and its associated metabolic abnormalities by administration of the pharmaceutical composition according to the present invention. It refers to all actions that are beneficially changed, and “improvement” refers to all actions that reduce parameters related to the desired disease, such as the degree of symptoms, by administering the composition according to the present invention.
  • the present invention provides a drug delivery kit containing the porphyrin-liposome; and/or a kit for preventing or treating cancer containing a porphyrin-liposome encapsulated with an anticancer agent.
  • the "kit” is not limited to the specific form or type, and any kit commonly used in the art can be used, and can carry out the purpose of the present invention in addition to the porphyrin-liposome of the present invention. It may further include any necessary components. For example, in addition to the porphyrin-liposome of the present invention, it may further include a storage solution, administration tool, drug, suspension for administration, etc., and may further include an instruction sheet containing information about the porphyrin-liposome of the present invention.
  • the present invention includes the steps of (S1) dissolving porphyrin and phospholipid in a first organic solvent; and
  • step (S2) It provides a method for producing the ultrasonic-sensitive porphyrin-liposome, comprising the step of hydrating the mixed solution prepared in step (S1) in an aqueous solution and stirring it.
  • the first organic solvent may be one or more selected from the group consisting of dimethylacetamide, dimethylformamide, dimethyl sulfoxide, chloroform, methanol, ethanol, and ether, but is not limited thereto.
  • the first organic solvent is ethanol.
  • porphyrin and phospholipid are as described above.
  • the step (S1) may involve dissolving other lipids in addition to porphyrin and DOPE in the first organic solvent.
  • the lipid may preferably be selected from DSPC, DSPE-mPEG2000, cholesterol, etc.
  • the mixing ratio of the above lipids including DOPE is as described above. For example, it may be included in a weight ratio of 1:1 to 10:0.1 to 1:2 to 20, but is not limited thereto.
  • the DSPC is 1 to 30% by weight compared to the total ethanol solution
  • the DSPE-mPEG2000 is 5 to 40% by weight compared to the total ethanol solution
  • the cholesterol is 1 to 15% by weight relative to the total ethanol solution
  • the DOPE may be dissolved in an amount of 20 to 80% by weight based on the total ethanol solution, but is not limited thereto.
  • the weight ratio of porphyrin:total lipid in the ethanol solution is 1:20 to 90, 1:20 to 80, 1:20 to 70, 1:20 to 60, 1:20 to 50, It may be 1:30 to 80, 1:40 to 80, 1:30 to 70, 1:30 to 60, 1:40 to 60, or 1:40 to 50, but is not limited thereto.
  • the first organic solvent in step (S1) may be one to which polysorbate is added, but is not limited thereto.
  • the aqueous solution may be ammonium sulfate, ammonium citrate, ammonium sulfate, or TEA-SOS, but is not limited thereto, and may be appropriately selected depending on the type of drug carried in the porphyrin-liposome. . Most preferably, the aqueous solution is ammonium sulfate.
  • step (S1) is performed at a temperature of 20 to 80°C, 30 to 80°C, 40 to 80°C, 50 to 80°C, 60 to 80°C, 65 to 80°C, 60 to 75°C, or 65 to 75°C. It may be performed in, but is not limited to this.
  • the production method may further include, but is not limited to, the step of evaporating the organic solvent after step (S2).
  • the production method may further include the step of extruding the hydrated porphyrin-liposome through an extruder after step (S2).
  • the temperature for extrusion of the porphyrin-liposome can be adjusted variously from room temperature to the phase transition temperature range of each material, and the number of extrusion can be repeated an appropriate number of times to make the size of the porphyrin-liposome uniform.
  • porphyrin-liposome preparation prepared according to the production method of the present invention can be used for drug delivery to injections, transdermal, nasal cavity, and lungs.
  • the technology and pharmaceutically appropriate carriers, additives, etc. required for such formulations are widely known to those skilled in the art, and in this regard, Remington's Pharmaceutical Sciences (19th ed., 1995) may be referred to. .
  • the concentration of phospholipids dissolved in the first organic solvent is 10 to 200 mg/mL, 10 to 150 mg/mL, 10 to 100 mg/mL, 10 to 80 mg/mL, and 10 to 60 mg/mL.
  • step (S2) is performed at a temperature of 20 to 80°C, 20 to 70°C, 20 to 65°C, 30 to 80°C, 40 to 80°C, 50 to 80°C, 40 to 70°C, or 50 to 70°C. It may be performed in, but is not limited to this.
  • the manufacturing method may further include adding and mixing a drug to the porphyrin-liposome solution after step (S2).
  • the mixing ratio of the porphyrin-liposome:drug is 20 to 1:1, 15 to 1:1, 10 to 1:1, 20 to 5:1, 15 to 5:1, 10 to 5:1, 9 to 5:1. It may be 1, or a weight ratio (w/w%) of 9 to 7:1, but is not limited thereto.
  • the mixing may occur for 30 minutes to 5 hours, 30 minutes to 4 hours, 30 minutes to 3 hours, 1 hour to 5 hours, 1 hour to 4 hours, or 1 hour to 3 hours; It may be performed at 100 to 1,000 rpm, 100 to 800 rpm, 100 to 500 rpm, 100 to 400 rpm, 100 to 350 rpm, or 100 to 300 rpm, but is not limited thereto. Additionally, the mixing may be performed at a temperature of 20 to 50°C, 20 to 45°C, 20 to 40°C, 30 to 50°C, 35 to 45°C, or 35 to 40°C, but is not limited thereto.
  • the term "combination thereof" included in the Markushi format expression means a mixture or combination of one or more selected from the group consisting of the components described in the Markushi format expression, It means containing one or more selected from the group consisting of constituent elements.
  • DOX Doxorubicin DOX-liposome or DLL Liposome loaded with doxorubicin DOX-porphyrin-liposome; or DLPL Liposome loaded with doxorubicin and porphyrin US Ultrasound irradiation group
  • DSPC 32.1 mg of DSPC, 57.0 mg of DSPE-mPEG2000, 23.5 mg of cholesterol, 206.6 mg of DOPE, and 3.99 mg of porphyrin were dissolved in ethanol at a total concentration of 256 mg/ml at 70°C.
  • the lipid solution dissolved in a completely transparent solution was mixed with a 250mM aqueous ammonium sulfate solution, and the lipid concentration at this time was maintained at 32mg/mL and the temperature at 60°C. Stirring of the mixed solution was performed for 1 hour to hydrate the phospholipid membrane, and the mixed lipid solution was homogenized using a high-temperature extruder (LIPEX, Evonic).
  • LIPEX high-temperature extruder
  • a polycarbonate filter was manufactured by gradually reducing the size from 0.8 ⁇ m-pore to 0.4 ⁇ m-pore size, and then to 0.2 ⁇ m-pore size. The process was repeated until the size of the prepared porphyrin-liposome reached 100-200 nm.
  • the particle size distribution of porphyrin-liposomes was measured by diluting porphyrin-liposomes 10 times and using Zetasizer Nano ZS (Malvern) using Dynamic Light Scattering (DLS) analysis.
  • the porphyrin-liposome solution whose particle size distribution was controlled by extrusion, was introduced into a dialysis membrane with a molecular weight (MWCO) of 15,000 Dalton and dialyzed in a 10% sucrose, 10mM L-histidine solution for 12 hours.
  • MWCO molecular weight
  • doxorubicin was added to the dialyzed porphyrin-liposome solution and stirred at 200-300 rpm for 2 hours at 37°C. Porphyrin-liposome and doxorubicin were mixed at a weight ratio of 8:1. After stirring for 2 hours, unencapsulated doxorubicin was separated using size exclusive chromatography (SEC). Encapsulated and unencapsulated doxorubicin were quantified by absorbance measurement at a wavelength of 480 nm.
  • the ultrasound-sensitive porphyrin-liposome prepared according to Example 1 and a general liposome were prepared and compared.
  • the diameter of the ultrasound-sensitive porphyrin-liposome of the present invention was measured to be 102 ⁇ 14 nm, and the diameter of the regular liposome was measured to be 103 ⁇ 11 nm. As a result, it was confirmed that there was no significant difference in the size distribution of the two. Subsequently, in order to confirm the shape of each liposome particle, it was photographed and compared with a TEM (Transmission Electron Microscope), and it was confirmed that there was no significant difference in shape or size (Figure 1).
  • TEM Transmission Electron Microscope
  • the ultrasound sensitivity of the ultrasound-sensitive porphyrin-liposome encapsulated with doxorubicin and the general liposome were compared. Specifically, this analysis was performed using doxorubicin loaded at 0.1 mg/mL based on doxorubicin, and the doxorubicin loading rate of each liposome and the drug release rate according to ultrasound irradiation were confirmed.
  • doxorubicin-encapsulated porphyrin-liposomes or general liposomes were loaded into a 24 well plate, and then the plate was subjected to 1.5 W/w at the bottom of the plate using an ultrasonic device (SP100, SONIDEL). cm 2 , 50%, and 10 minutes.
  • SP100, SONIDEL ultrasonic device
  • cm 2 50%, and 10 minutes.
  • doxorubicin released from the liposome and the liposome from which doxorubicin was released were separated using size exclusive chromatography (SEC). Quantitative analysis of doxorubicin was performed by measuring absorbance at 480 nm. The results are shown in Figure 2 and Table 2.
  • the drug encapsulation rate (95.8%) of the ultrasonic liposome of the present invention was found to be comparable to that of general liposomes (97.1%).
  • the rate of doxorubicin released from the liposomes was only less than 5% even when irradiated with ultrasound, but in the case of the porphyrin-liposome according to the present invention, the drug release rate was more than 95% when irradiated with ultrasound, and most of the porphyrin -It was confirmed that the liposome responded to ultrasound and released the drug.
  • MTT analysis was performed using 4T1 cells, a cancer cell line.
  • the anticancer effects of doxorubicin-encapsulated porphyrin-liposome, doxorubicin-encapsulated regular liposome (does not contain porphyrin), and free doxorubicin (free DOX) were compared, and each group was divided into an ultrasound irradiation group and an ultrasound untreated group.
  • the 4T1 cell line was appropriately dispensed into each well of a 96-well plate and cultured overnight.
  • Free doxorubicin, ultrasound-irradiated liposomes (+US), or non-ultrasounded liposomes (-US) were added to the cultured cells in the designated wells. Each was processed. Each liposome or free doxorubicin was treated at a concentration of 2 to 10 ⁇ g/mL. Afterwards, all added drugs were removed, the cells were washed, and the degree of proliferation of cancer cells was compared while further cultured with cell culture medium to which MTT was added.
  • the experimental groups were divided into the following: (i) negative control administered saline, (ii) free doxorubicin (Free DOX) without sonication, (iii) DOX-liposome (LIP-DOX) without sonication, ( iv) DOX-Porphyrin-Liposome without sonication (PPR-LIP-DOX), (v) ultrasound treatment only, (vi) free doxorubicin with ultrasound pretreatment (Free DOX+US), (vii) ultrasound pretreatment DOX-liposomes (LIP-DOX+US), and (viii) ultrasound-pretreated DOX-porphyrin-liposomes (PPR-LIP-DOX+US).
  • Each substance was administered a single intravenous injection at a dose of 2 mg/kg (volume 200 ⁇ L) based on doxorubicin, and the mice were monitored for 2 weeks after injection, with body weight and condition recorded on days 3, 6, 9, 12, and 15 after injection. recorded.
  • the ultrasound-sensitive porphyrin-liposome according to the present invention can maintain structural stability even when encapsulating a drug, minimizing the risk of non-specific drug release, and has excellent sensitivity to ultrasound. It was confirmed that release could be appropriately controlled (Figure 5).
  • porphyrin-liposomes loaded with anticancer drugs demonstrated better anticancer effects than free anticancer drugs, confirming that they had excellent drug delivery efficiency. Therefore, the ultrasound-sensitive porphyrin-liposome according to the present invention can further improve the delivery efficiency and pharmacological effect of the encapsulated drug, and is expected to be useful in the field of treatment of various diseases and drug delivery.
  • the present invention relates to ultrasound-sensitive porphyrin-liposomes capable of releasing drugs in response to ultrasound.
  • the ultrasound-sensitive porphyrin-liposome of the present invention is characterized in that it contains porphyrin. Porphyrin exposed to ultrasound reacts with water to generate reactive oxygen species (Reactive Oxygen Species) and constitutes a lipid bilayer of the porphyrin-liposome. Phospholipids react to reactive oxygen species generated by porphyrin, causing lipid peroxidation, which can eventually cause the structure of the lipid bilayer to collapse and release the encapsulated drug.
  • Reactive Oxygen Species reactive oxygen species
  • Phospholipids react to reactive oxygen species generated by porphyrin, causing lipid peroxidation, which can eventually cause the structure of the lipid bilayer to collapse and release the encapsulated drug.
  • the ultrasound-sensitive porphyrin-liposome according to the present invention can appropriately control drug release using ultrasound, and also maintains excellent structural stability even when the drug is encapsulated, thereby minimizing non-specific drug delivery.
  • the porphyrin-liposome of the present invention can improve the drug delivery efficiency and pharmacological effect of the drug, and can be used as an excellent drug carrier for various diseases by carrying various drugs.

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Abstract

La présente invention concerne des liposomes porphyriniques sensibles aux ultrasons capables de libérer des médicaments en réponse à des ultrasons. Les liposomes porphyriniques sensibles aux ultrasons selon la présente invention contiennent de la porphyrine. Lorsqu'elle est exposée aux ultrasons, la porphyrine réagit avec l'eau pour générer des espèces réactives de l'oxygène. Les phospholipides constituant la bicouche lipidique des liposomes porphyriniques réagissent avec les espèces réactives de l'oxygène produites par la porphyrine pour induire la peroxydation lipidique, ce qui conduit par conséquent à l'affaissement de la structure de bicouche lipidique, permettant aux médicaments enfermés d'être libérés.
PCT/KR2023/010440 2022-07-22 2023-07-19 Liposome porphyrinique sensible aux ultrasons et son utilisation WO2024019542A1 (fr)

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JP2017105805A (ja) * 2009-10-16 2017-06-15 ユニバーシティ・ヘルス・ネットワーク ポルフィリンナノ小胞
KR20180120671A (ko) * 2015-11-23 2018-11-06 유니버시티 오브 얼스터 초음파역학 치료를 위한 미세거품-화학치료제 복합체
KR20190001022A (ko) * 2017-06-26 2019-01-04 서강대학교산학협력단 광감작제를 포함한 마이크로버블-나노입자 복합체 및 이를 포함하는 항암 치료제

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JP2017105805A (ja) * 2009-10-16 2017-06-15 ユニバーシティ・ヘルス・ネットワーク ポルフィリンナノ小胞
KR20180120671A (ko) * 2015-11-23 2018-11-06 유니버시티 오브 얼스터 초음파역학 치료를 위한 미세거품-화학치료제 복합체
KR20190001022A (ko) * 2017-06-26 2019-01-04 서강대학교산학협력단 광감작제를 포함한 마이크로버블-나노입자 복합체 및 이를 포함하는 항암 치료제

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